Programmed cell death in the plant immune system.

Cell death has a central role in innate immune responses in both plants and animals. Besides sharing striking convergences and similarities in the overall evolutionary organization of their innate immune systems, both plants and animals can respond to infection and pathogen recognition with programmed cell death. The fact that plant and animal pathogens have evolved strategies to subvert specific cell death modalities emphasizes the essential role of cell death during immune responses. The hypersensitive response (HR) cell death in plants displays morphological features, molecular architectures and mechanisms reminiscent of different inflammatory cell death types in animals (pyroptosis and necroptosis). In this review, we describe the molecular pathways leading to cell death during innate immune responses. Additionally, we present recently discovered caspase and caspase-like networks regulating cell death that have revealed fascinating analogies between cell death control across both kingdoms.

Differential induction of the Arabidopsis thaliana Thi2.1 gene by Fusarium oxysporum f. sp. matthiolae.

The Arabidopsis thaliana Thi2.1 gene is inducible by necrotrophic fungi through a signal transduction pathway different from that for pathogenesis-related (PR) proteins. We have identified three ecotypes that are susceptible (Col-2, Ler, and Ws) and two ecotypes that are resistant (Mt-0 and Uk-4) to spray inoculation with Fusarium oxysporum f. sp. matthiolae. The Thi2.1 transcript level after infection correlates with resistance, being 5 to 10 times higher in the resistant than in the susceptible ecotypes. The beta-glucuronidase (GUS) expression of a Thi2.1-promoter-uidA fusion (with a promoter derived from Col-2) is on the average almost 10 times higher in the Uk-4 background than in the Col-2 background. This confirms the results obtained by Northern (RNA) blots and indicates that Uk-4, and probably other resistant ecotypes too, might have a more sensitive recognition system for F. oxysporum f. sp. matthiolae or might have a signal transduction system that gives a higher amplification of the original recognition signal. Our results suggest a role of the Thi2.1 gene in resistance against F. oxysporum f. sp. matthiolae and perhaps other necrotrophic fungi.

Overexpression of an endogenous thionin enhances resistance of Arabidopsis against Fusarium oxysporum.

Thionins are antimicrobial proteins that are thought to be involved in plant defense. Concordant with this view, we have recently shown that the Arabidopsis thionin Thi2.1 gene is inducible by phytopathogenic fungi. Here, we demonstrate that constitutive overexpression of this thionin enhances the resistance of the susceptible ecotype Columbia (Col-2) against attack by Fusarium oxysporum f sp matthiolae. Transgenic lines had a reduced loss of chlorophyll after inoculation and supported significantly less fungal growth on the cotyledons, as evaluated by trypan blue staining. Moreover, fungi on cotyledons of transgenic lines had more hyphae with growth anomalies, including hyperbranching, than on cotyledons of the parental line. No transcripts for pathogenesis-related PR-1, PR-5, or the pathogen-inducible plant defensin Pdf1.2 could be detected in uninoculated transgenic seedlings, indicating that all of the observed effects of the overexpressing lines are most likely the result of the toxicity of the THI2.1 thionin. Our findings strongly support the view that thionins are defense proteins.

ESTs reveal a multigene family for plant defensins in Arabidopsis thaliana.

Plant defensins, formerly named gamma-thionins, are a group of small, cysteine-rich, basic, and antimicrobial plant proteins. Random sequencing of expressed sequence tags (ESTs) in Arabidopsis thaliana has revealed several different plant defensin genes in this plant species which can be grouped into two subfamilies. We have used one EST of each subfamily to study the expression of the corresponding genes in A. thaliana. Pdf2.3 is constitutively expressed in seedlings, rosettes, flowers, and siliques and is not inducible in seedlings either by methyl jasmonate, salicylate, ethephon, and silver nitrate or by several different phytopathogenic fungi. The expression of a second gene, Pdf1.2, is in untreated plants only detectable in rosettes. In seedlings, it is inducible by methyl jasmonate, silver nitrate, and different phytopathogenic fungi, notably Fusarium oxysporum f. sp. matthiolae. The regulation of Pdf1.2 resembles that of the pathogen-inducible thionin gene Thi2.1.

The sink-specific and stress-regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors, and pathogen challenge.

A cDNA for the Arabidopsis STP4 gene (for sugar transport protein 4) was isolated, and the properties of the encoded protein were studied in Schizosaccharomyces pombe. The STP4 monosaccharide H+ symporter is composed of 514 amino acids and has a calculated molecular mass of 57.1 kD. RNA gel blot analyses revealed that STP4 is expressed primarily in roots and flowers of Arabidopsis. This was shown in more detail with STP4 promoter-beta-glucuronidase (GUS) plants yielding strong STP4-driven GUS activity in root tips and anthers. Wounding of plants transformed with STP4-GUS constructs resulted in a rapid increase in GUS activity in cells directly adjacent to the lesion. This was confirmed by RNase protection analyses in Arabidopsis wild-type plants showing a strong, wound-induced increase in STP4 mRNA levels. STP4 expression was induced rapidly in suspension-cultured Arabidopsis cells that were treated with the Pseudomonas syringae elicitor or with chitin or in Arabidopsis plants that were exposed to fungal attacks. Our data suggest that the role of STP4 is to catalyze monosaccharide import into classic sinks, such as root tips and anthers, and, most importantly, to meet the increased carbohydrate demand of cells responding to environmental stress.

Cytosolic and plastidic chorismate mutase isozymes from Arabidopsis thaliana: molecular characterization and enzymatic properties.

The three aromatic amino acids phenylalanine, tyrosine, and tryptophan are synthesized in the plastids of higher plants. There is, however, biochemical evidence that a cytosolic isoform exists of the enzyme catalysing the first step of that branch of the pathway which is specific for the synthesis of phenylalanine and tyrosine, i.e. chorismate mutase (CM). We now report on the isolation of a cDNA clone encoding a cytosolic CM isozyme from Arabidopsis thaliana that was identified by complementing a CM-deficient Escherichia coli strain. The deduced amino acid sequence of this isozyme was 50% identical to that of a previously isolated plastidic CM, and 41% identical to that of yeast CM. The organ-specific expression patterns of the two CM genes were rather similar, but only the gene encoding the plastidic isozyme was elicitor- and pathogen-inducible. The plastidic CM expressed in E. coli was activated by tryptophan and inhibited by phenylalanine and tyrosine, whereas the cytosolic isozyme was insensitive. The existence of a cytosolic CM isozyme implies that either a cytosolic pathway (partial or complete) for the biosynthesis of phenylalanine and tyrosine exists, or that prephenate, originating from chorismate in the cytosol, is utilized for the synthesis of metabolites other than these two aromatic amino acids.

The evaluation of screening methods to identify medically unnecessary hospital stay for patients with pneumonia.

OBJECTIVES: This study compared three methods to screen charts of pneumonia patients for excess days. METHODS: A derivation data set was used to statistically derive a severity measure to predict length of stay for pneumonia patients and to refine a clinical algorithm for identifying excess stay. A validation data set was used to compare three computerized methods to screen for unnecessary hospital days: (1) an observed length of stay greater than a target value; (2) an observed stay greater than predicted for the specific patient; and (3) an algorithm that tested whether there were clinical justifications for the entire hospital stay. RESULTS: The sensitivity and specificity for detecting excess stay for the three methods were (1) 0.48 and 0.85 for the observed stay greater than the target value; (2) 0.56 and 0.73 for observed stay greater than predicted; and (3) 0.83 and 0.85 for the algorithm. CONCLUSIONS: These results suggest that computerized clinical algorithms may provide a useful method to detect unnecessary hospital stay.

An Arabidopsis thaliana thionin gene is inducible via a signal transduction pathway different from that for pathogenesis-related proteins.

Two cDNAs encoding thionin preproteins have been isolated from Arabidopsis thaliana. The corresponding genes have been designated Thi2.1 and Thi2.2. Southern blot analysis suggests that A. thaliana most probably contains single genes for both thionins. Thi2.2 transcripts have a low basal level in seedlings and show circadian variation. Thi2.2 transcripts were also detected in rosette leaves. No potent elicitors have been found for the Thi2.2 gene. Transcripts of the Thi2.1 gene are not detectable in seedlings but are present in rosette leaves and at a very high level in flowers and in siliques. The expression of the Thi2.1 gene is highly inducible in seedlings by pathogens, silver nitrate, and methyl jasmonate, but not by salicylate, indicating that the gene is induced by a signal transduction pathway that is at least partly different from that for the pathogenesis-related proteins.

Legionella pneumophila serogroup Lansing 3 isolated from a patient with fatal pneumonia, and descriptions of L. pneumophila subsp. pneumophila subsp. nov., L. pneumophila subsp. fraseri subsp. nov., and L. pneumophila subsp. pascullei subsp. nov.

Previous DNA relatedness and enzyme electrophoretic mobility studies indicated heterogeneity among strains of Legionella pneumophila serogroups 1, 4, 5, and Lansing 3 (a new, as yet unnumbered serogroup). In this study 60 L. pneumophila strains were studied by DNA hybridization (hydroxyapatite method) to assess their genomic relatedness. These strains were also studied biochemically and serologically to determine whether they formed one or more phenotypic groups. DNA relatedness studies identified three groups. DNA group 1 contained the type strain Philadelphia 1 and strains from serogroups 1 through 14 of L. pneumophila. The average relatedness of DNA group 1 strains was 88% at 60 degrees C with 1.1% divergence in related sequences and 85% at 75 degrees C. DNA group 2 contained strain Los Angeles 1, the reference strain of serogroup 4, and strains of serogroups 1, 4, 5, and Lansing 3, an unnumbered serogroup. Average relatedness of DNA group 2 strains was 84% at 60 degrees C with 0.7% divergence and 87% at 75 degrees C. Reciprocal relatedness of DNA groups 1 and 2 was approximately 67% at 60 degrees C with 6.0% divergence and 48% at 75 degrees C. DNA group 3 strains were in serogroup 5. They were 98% related at 60 degrees C with 0.5% divergence and 97% related at 75 degrees C. Reciprocal relatedness of DNA group 3 and DNA group 1 was approximately 74% at 60 degrees C with 5.3% divergence and 43% at 75 degrees C, and reciprocal relatedness of DNA groups 3 and 2 was 66% at 60 degrees C with 5.7% divergence and 55% at 75 degrees C. The DNA groups could not be separated biochemically or serologically or by cell wall fatty acid and isoprenoid quinone composition. Three subspecies of L. pneumophila are proposed to accommodate the three DNA groups: L. pneumophila subsp. pneumophila subsp. nov. for DNA group 1, L. pneumophila subsp. fraseri subsp. nov. for DNA group 2, and pneumophila subsp. pascullei subsp. nov. for DNA group 3.

Progressive reorganization of the host cell cytoskeleton during adenovirus infection.

Infection of cells with adenovirus lead to a characteristic reorganization of all cytoskeleton systems, starting with alterations at the microtubuli of the cells. During this progress, the flat, extended, and polar morphology of the cytoskeleton became nonpolar and rounder. These rearrangements were initiated before the appearance of adenovirus structural proteins hexon and fiber, as well as before the shutoff of host protein synthesis. We conclude that these alterations reflect a specific reorganization rather than an unorganized breakdown of the cell during adenovirus infection.